FSCB基因敲除对雄性小鼠生殖功能的影响  

Knockout of fibrous sheath CABYR-binding on reproductive function in male mice

作　　者：周庭友 张勇[1] 张军[1] 刘旭东[1] 李珂[1] 周波[1] 靳风烁[1] 李彦锋[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所泌尿外科,重庆400042

出　　处：《第三军医大学学报》2016年第15期1729-1734,共6页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81170617)~~

摘　　要：目的利用fscb基因敲除小鼠,探讨FSCB蛋白在小鼠精子运动和授精功能中的作用。方法应用免疫荧光观察fscb基因敲除雄鼠睾丸组织及精子中FSCB蛋白表达情况;应用计算机辅助精液分析(computer assisted sperm analysis,CASA)系统分析fscb基因敲除小鼠精子在体外的运动特征、活率和运动参数改变情况;在体外获能培养条件下检测精子的体外受精能力;将fscb基因敲除纯合子(KO)、杂合子(HET)、野生型(WT)雄鼠分别与野生型C57BL/6J成年雌鼠交配,观察雌鼠怀孕率、怀孕胚胎数量、自然产仔数及子代成年雄鼠生殖腺形态学是否存在差异。结果免疫荧光显示纯合子雄鼠睾丸组织及精子中均无FSCB蛋白表达,杂合子雄鼠睾丸组织及精子中FSCB蛋白表达与野生型相比明显减低。CASA显示各组雄鼠精子数、活率、各项运动参数差异均无统计学意义(P>0.05);野生型、杂合子和纯合子雄鼠精子与透明带完整卵细胞及与无透明带卵细胞结合的精子平均数、卵细胞的受精率差异均无统计学意义(P>0.05);各组雄鼠对应雌鼠的受孕率差异无统计学意义(P>0.05);野生型、杂合子和纯合子对应雌鼠受孕后每只雌鼠胚胎数分别为(7.00±1.41)、(7.33±1.53)、(7.20±1.48)只,每窝自然产仔数分别为(6.00±2.00)、(6.75±1.71)、(7.80±1.30)只,组间比较差异无统计学意义(P>0.05);各组子代成年雄鼠生殖腺发育差异无统计学意义(P>0.05)。结论敲除fscb基因对雄性小鼠精子的运动功能及体内外受精能力均无明显影响,提示该基因所编码的FSCB蛋白的功能对于精子的运动和授精能力的维持可能是非必需的或可被替代的。Objective To investigate the role of fibrous sheath CABYR-binding （FSCB） protein in the sperm motility and fertility in fscb gene knockout male mice. Methods In fscb-gene knockout mice the expression of FSCB protein in testicular tissue and sperm was observed by immunohistochemistry. In vitro the sperm motility, viability and corresponding parameters were analyzed by computer-aided sperm analysis （CASA）. In vitro the fertilization ability of sperm from Fscb-gene knock out mice was studied. Male mice of homozygous Fscb-gene knock out（KO）, heterozygote（Her） and wild type（WT） were mated with C57BL/6J adult female WT mice. The pregnancy rate, and numbers of the embryos and natural born offspring were counted. Morphological changes of the gonads were also observed in filial generation of male mice. Results The immune fluorescence images detected no expression of FSCB protein in the testicular tissue and sperm of KO mice. The expression of FSCB protein in the testicular tissue and sperm of Het mice was significantly lower than that in WT mice. The amount and motility of sperm and the related motility parameters were not statistically different among the groups （ P 〉 0.05 ）. Furthermore, the data of in vitro fertilization （IVF） experiment showed that the sperm fertility in KO, Het and WT groups was not widely divergent from each other. The average number of sperms binding to eggs with intact zona pellucida and zona-free eggs, the fertilization rate of eggs and the pregnancy rate of the female mice was not significantly different （P 〉 0.05 ）. Specifically, the number of embryos for each female in WT, Het and KO groups were 7.00 ± 1.41, 7.33 ± 1.53, and 7.20 ± 1.48, accordingly, the number of natural born offsprings was 6.00 ± 2.00, 6.75 ± 1.71, and 7.80 ± 1.30. In addition, the gonado genesisin filial generation of adult male mice had no obvious difference. Gonclusion Fscb gene knockout does not affect the ability of sperm motility and fertilization in vitro and in vivo, suggesti

关 键 词：FSCB 基因敲除 精子 生殖力 

分 类 号：R339.21[医药卫生—人体生理学] R341[医药卫生—基础医学]